Trehalulose-containing composition, its preparation and use

ABSTRACT

A trehalulose-containing composition, its preparation and use.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a divisional of prior U.S. patent applicationSer. No. 13/263,361, filed Dec. 6, 2011, by Wolfgang Wach, Thomas Rose,Michael Klingeberg, Siegfried Peters, Tillmann Dörr Stephan Theis, JörgKowalczyk and Stephan Hausmanns, entitled “TREHALULOSE-CONTAININGCOMPOSITION, ITS PREPARATION AND USE,” the entire contents of which areincorporated herein by reference. U.S. patent application Ser. No.13/263,361 is a 35 U.S.C. §371 National Phase conversion ofPCT/EP2010/002308, filed Apr. 15, 2010, which claims the benefit of U.S.patent application Ser. No. 12/386,187, filed Apr. 15, 2009, abandoned,the entire contents of which are incorporated herein by reference.

TECHNICAL FIELD

The present invention relates to a trehalulose-containing composition,that means vitalose, a process for its preparation and methods for theuse thereof.

BACKGROUND

Trehalulose (1-O-α-D-glucopyranosyl-D-fructose) and isomaltulose(palatinose, 6-O-α-D-glucopyranosyl-D-fructose) are known structuralisomers of sucrose. These isomers are naturally found in honey in smallquantities. Both of them are known as non-cariogenic sugars. However,their physical and chemical characteristics are somewhat different, inparticular in view of the fact that trehalulose is more soluble in waterthan isomaltulose with the consequence that their use as foodingredients is different.

Both, isomaltulose and trehalulose, are also known from for instance EP1 424 074 A1 as being heterodisaccharides, which in comparison tosucrose, show a reduced hydrolysis rate in the small intestine of humanor animal consumers, thereby suggesting the provision of nutritionalcompositions for controlling blood sugar levels, in particular for theuse in patients suffering from diabetes and glucose intolerance, or forobesity prevention. However, the document is silent on the specific roleand relevance of isomaltulose and trehalulose contained in thenutritional compositions.

EP 0 483 755 B1 discloses processes to prepare compositions comprisingtrehalulose and isomaltulose. In particular, Pseudomonas mesoacidophilaMX-45 (FERM-BP 3619) or Agrobacterium radiobacter MX-232 (FERM-BP 3620)was used to convert a sucrose containing medium into a primarilytrehalulose, but also significant amounts of isomaltulose containingsolution. Thus, the disclosed methods make use of the capability of theα-glucosyltransferase of either Pseudomonas mesoacidophila orAgrobacterium radiobacter to convert sucrose both into trehalulose andisomaltulose, wherein predominantly trehalulose is obtained.

EP 0 794 259 B1 discloses the preparation of a trehalulose-containingpolysaccharide composition using a maltose/trehalose converting enzymefrom Pseudomonas putida, Thermus ruber, Thermus aquatica or Pimelobacterto produce the trehalulose-containing composition from a sucrosesolution.

EP 0 091 063 A2 discloses a process for the preparation of isomaltuloseusing immobilized bacterial cells, in particular cells fromProtaminobacter rubrum (CBS 574.77). The process disclosed therein usesa sucrose-containing solution, which is subjected to these immobilizedcells so as to obtain a primarily isomaltulose containing composition,which, however, also comprises trehalulose.

EP 1 393 637 A1 discloses agents for sustaining concentration andattentiveness, in particular food and drinks containing said agents. Thedocument discloses products for human consumption, which are said toimprove the concentration and attention due to the presence ofisomaltulose contained in the product.

EP 0 983 374 B1 discloses processes for the simultaneous production ofisomaltulose and/or trehalulose and betain.

All of these processes aim to produce either a trehalulose or aisomaltulose-containing composition or even pure trehalulose orisomaltulose so as to make use of the known physiological properties, inparticular their non-cariogenity and their slow hydrolyses rate in thesmall intestine. Although isomaltulose is an attractive agent fordeveloping products suitable to prevent and treat overweight, obesity,diabetes and other glucose or insulin metabolism-related conditions anddiseases, there still remains the need to provide even more effectiveactive ingredients.

The production of trehalulose-containing compositions, also termedvitalose, which comprise a high content of isomaltulose is for a numberof applications, such as jelly or fruit juices, not desirable, sinceisomaltulose tends to crystallize out, in particular in applications inan aqueous surrounding. Although the preparation of compositionscomprising a high content of trehalulose is for some applicationsadvantageous, it has for other applications the disadvantage of itsseverely inhibited crystallization capability. Up to now there has beenno report on processes to obtain trehalulose-containing compositions,that means vitalose, which comprise both trehalulose and isomaltulose insignificant amounts, and which allow the production thereof in an easy,inexpensive and commercially suitable way and which provide theadvantage of its composition, in particular its ratio of trehalulose toisomaltulose, being adjustable according to the specific needs of theparticular application of the produced trehalulose-containingcomposition, that means vitalose. Furthermore, there is the need in theart to provide improved means and methods for the prophylaxes andtreatment of conditions and diseases related to the glucose and/orinsulin metabolism in the human or animal body, in particular forspecific groups of patients, which up to now have not been considered tobe suitable for being supplied with a carbohydrate, in particularsugar-containing diet.

SUMMARY

Thus, the technical problem underlying the present invention is toprovide improved trehalulose- and isomaltulose-containing compositions,in the following termed “vitalose” or “trehalulose-containingcompositions”, which overcome the above-identified disadvantages andwhich in particular enable advantageous applications, in particular inthe prophylaxes and therapy of conditions and diseases related to theglucose and/or insulin metabolism, preferably for specific groups ofconsumers or patients. Thus, in the context of the present invention theterms “trehalulose-containing composition” and “vitalose” refer to thesame trehalulose- and isomaltulose-containing composition and are usedinterchangeably. The technical problem underlying the present inventionis also to provide processes for the production oftrehalulose-containing compositions, that means vitalose, in anindustrial scale, in particular processes which allow their productionin a particularly variable way, i.e. in a way allowing the productionof, preferably a preselected, product composition, preferably exhibitinga specific, preferably preselected trehalulose/isomaltulose ratio.

The present invention solves its problem by the teaching of theindependent claims.

Thus, in a preferred embodiment the present invention solves itstechnical problem by providing a process for the preparation of atrehalulose-containing composition, that means vitalose, wherein (a) asucrose containing composition is contacted under appropriate conditionswith cells or cell extracts from microorganisms of the genus Pseudomonasand the genus Protaminobacter and (b) the trehalulose-containingcomposition, that means vitalose, is produced.

Thus, the above-identified process of the present invention foreseessubjecting a sucrose-containing composition, in particular an aqueousmedium comprising sucrose, to an enzymatic activity, in particular anα-glucosyltransferase activity, preferably contained in a cell or cellextract, from microorganism of the genera Pseudomonas andProtaminobacter, for a time period and under conditions suitable toconvert the sucrose into a trehalulose-containing composition, thatmeans vitalose. In a preferred embodiment the trehalulose-containingcomposition, that means vitalose, is isolated and optionally furtherpurified, preferably subsequent to the complete, i.e. 100% conversion,or almost complete, that means at least a 95, 96, 97, 98 or 99%conversion of the sucrose. The obtained trehalulose-containingcomposition, that means vitalose, may be further purified according toprior art methods and according to the particular needs resulting fromthe intended application. In a preferred embodiment it is for instancepossible to further purify the obtained trehalulose-containingcomposition, that means vitalose, by chromatography, filtration,deionisation, decolouration, enzymatic treatment, catalytic treatmentand/or fractionation.

Thus, the present invention provides the advantageous and unexpectedteaching that cells or cell extracts from a microorganism of the genusPseudomonas and cells or cell extracts from a microorganism of the genusProtaminobacter can be used simultaneously and together in one singleprocess to generate from a sucrose-containing composition atrehalulose-containing composition, that means vitalose, preferably witha preselected and easily adaptable composition of its components.Favourably, the present teaching allows to produce atrehalulose-containing composition, that means vitalose, with a broadrange of ratios of its components, particularly trehalulose andisomaltulose. In particular, the combined and simultaneous use of cellsor cell extracts from both Pseudomonas and Protaminobacter produces anadvantageous composition, in particular sweetener composition,comprising mainly both of trehalulose and isomaltulose, in the followingtermed “trehalulose-containing composition”, that means vitalose, andwhich provides the advantages detailed below, namely which shows anunexpected glycemic and insulinemic behaviour and opens up furtheradvantageous methods and uses thereof, in particular in controlling bodyweight and in general glucose and insulin metabolism-related conditionsand diabetes. The trehalulose-containing composition, that meansvitalose, of the present invention is essentially non-cariogenic,preferably completely non-cariogenic, and has a low glycemic and lowinsulinemic index. The present composition is a low calorie composition,which is suitable for health oriented consumers and also in particularfor diabetics and combines a pleasant sweetening effect with goodbodying properties, which advantageously can be prepared in solid form,but also advantageously can be used in form of a syrup, in particular asuspension or in form of a solution. In particular, the composition ofthe present invention does not crystallize out in the concentrationsused. The compositions of the present invention have—without being boundby theory—a composition, attributed mainly by a specific ratio of itvarious components, in particular the ratio of trehalulose toisomaltulose and to the minor carbohydrate components isomaltose,isomelezitose and oligomers.

DETAILED DESCRIPTION

In the following, reference to percent (%) is a reference to weight-%,unless otherwise specified. In the context of the present invention, allamounts, in particular relative amounts of components of a composition,do not exceed 100%, preferably add up to 100% based either on the drymatter of the composition or the overall weight of the composition, asindicated.

In the context of the present invention, a sucrose-containingcomposition is preferably a composition comprising 1 to 100%, preferably1 to 99 weight-% or 90 to 100 weight-% sucrose (weight-% of dry matter).In a particularly preferred embodiment a sucrose-containing compositionhas a sucrose content of preferably 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30,40, 50, 60, 70 or 80 to 98 weight-%, preferably 3, 4, 5, 6, 7, 8, 9, 10,20, 30, 40, 50, 60, 70 or 80 to 97 weight-%, preferably 4, 5, 6, 7, 8,9, 10, 20, 30, 40, 50, 60, 70 or 80 to 96 weight-%, preferably 5, 6, 7,8, 9, 10, 20, 30, 40, 50, 60, 70 or 80 to 95, 96, 97, 98 or 99 weight-%,preferably 60 to 90 weight-%, preferably 70 to 80 weight-%, preferably30 to 60 weight-% or preferably 40 to 50 weight-% sucrose (each based ondry matter of the composition). Most preferably, the sucrose content ofthe sucrose-containing composition is from 90 to 99 weight-%, preferably90 to 98 weight-% (each based on dry matter of composition).

In a particularly preferred embodiment, the sucrose-containingcomposition is used in liquid form, particularly is dissolved orsuspended in an aqueous medium, preferably is an aqueoussucrose-containing solution or suspension. Preferably, theabove-identified sucrose-containing composition may be dissolved orsuspended in an aqueous medium, e.g. water, so as to obtain a solutionor suspension comprising 0.1 to 80 weight-%, preferably 40 to 75weight-%, preferably 40 to 60 weight-%, in particular 10 to 60 weight-%,preferably 20 to 55 weight-% of the sucrose-containing composition inwater, thus adding up with water or an aqueous medium to 100% (based onoverall weight of solution or suspension). In a particularly preferredembodiment the sucrose-containing solution or suspension has a sucrosecontent of 0.1 to 80 weight-%, preferably 5 to 30 weight-%, 20 to 30weight-%, 20 to 60 weight-%, 30 to 60 weight-% or 40 to 75 weight-%,preferably 40 to 60 weight-%, in particular 10 to 60 weight-%,preferably 20 to 55 weight-% sucrose (based on overall weight of theliquid solution or suspension). In a particularly preferred embodimentthe sucrose-containing solution or suspension may also be a thin juiceor thick juice from a sugar factory having preferably a dry substancecontent of 5 to 30%, preferably 20 to 27%. It is also possible to usemolasses or other impure sucrose compositions found in sugar factories.

In the context of the present invention a cell or cell extract from amicroorganism of the genus Pseudomonas is suitable under appropriateconditions to convert sucrose into a composition comprising trehaluloseand isomaltulose. In the context of the present invention a cell or cellextract from a microorganism of the genus Protaminobacter is suitableunder appropriate conditions to convert sucrose into a compositioncomprising trehalulose and isomaltulose. Thus, in the present inventionthe cells or cell extracts from the microorganism of the generaPseudomonas and Protaminobacter are cells or cell extracts capable offorming trehalulose and isomaltulose from sucrose.

In the context of the present invention a cell extract is meant to be anextract or a part from one or more of the cells of the microorganismused in accordance with the present invention, for instance refers todisrupted cells. In a particularly preferred embodiment the cell extractis also referring to a solution or suspension of an enzyme or enzymesystem having the capability to convert sucrose into trehalulose andisomaltulose, in particular is a solution or suspension containing theα-glucosyltransferase from Pseudomonas and/or Protaminobacter, thatmeans preferably an α-glucosyltransferase or sucrose-6-glucosylmutase(EC 5.4.99.10). Thus, the present invention foresees the use of cells orcell extracts, wherein the term “cell extract” in a preferred embodimentalso means enzymes, this term also including enzyme systems, comprisingsaid α-glucosyltransferase. According to the invention it is possible touse whole or disrupted cells of the microorganism with the cells actingas carrier for the enzyme system. The cells, in particular theimmobilized cells, are preferably dead. The enzyme or enzyme system usedaccording to the present invention can be extracted from cells from themicroorganism by conventional techniques, for example by solventextraction, French press, lyophilisation and/or enzymatic treatment. Itis possible to use auxiliary processes to facilitate extraction, such asdisrupting the cells or osmotic shocking. The extracted enzyme or enzymesystem may be purified further or may be used as it is. The presentinvention not only foresees the use of the naturally occurring wild typeenzymes, that means the α-glucosyltransferase, from Pseudomonas and/orProtaminobacter, but also foresees to use genetically engineeredderivatives thereof as well as cells from genetically engineered mutantsof Pseudomonas and/or Protaminobacter.

In a particularly preferred embodiment of the present invention, themicroorganism of the genus Pseudomonas is a microorganism of the speciesPseudomonas mesoacidophila, preferably is Pseudomonas mesoacidophilaMX-45, preferably P. mesacidophila MX-45 (FERM-BP 3619, said organism isdisclosed for instance in EP 0 483 755 B1 or EP 0 625 578 B1 depositedalso under FERM 11808) or also publically available from Nagai et al.(Biosc. Sci. Biotech. Biochem. 58 (10), (1994) 1789-1793)). In aparticularly preferred embodiment of the present invention themicroorganism of the genus Protaminobacter is a microorganism of thespecies Protaminobacter rubrum, preferably is Protaminobacter rubrum CBS574.77 (Protaminobacter rubrum CBS 574.77 is for instance described inEP 0 091 063 A2 or EP 0 625 578 B1).

The appropriate conditions for converting the sucrose-containingcomposition into the trehalulose-containing composition, that meansvitalose, are in a preferred embodiment a temperature of 10° C.,preferably 15° C. to 40° C., preferably 10° C. to 37° C., preferably 25°C. to 40° C., preferably 30° C. to 40° C., preferably 10° C. to 25° C.,preferably 15° C. to 30° C., 18° C. to 26° C. and most preferably 10° C.to 20° C., and particularly preferred 10 to 17° C.

In a furthermore preferred embodiment the process is carried out at a pHvalue from 5.0 to 9.0, preferably 5.0 to 7.0, preferably 6.0 to 7.0.

In a furthermore preferred embodiment the sucrose-containing compositionis subjected to the cells or cell extracts from the genera from themicroorganisms Protaminobacter and Pseudomonas for a time period andunder conditions suitable to achieve a conversion of sucrose to thetrehalulose-containing composition, that means vitalose, of 20 to 100%,preferably 30 to 95, 96, 97, 98, 99% and preferably 100%, preferably 40to 98%, preferably 50, 60 or 70 to 97%, preferably 50, 60, 70 or 80 to98%, preferably 75 to 98%, most preferably 50 to 99%, most preferably 60to 98%, 85 to 100% and preferably more than 95, 96, 97, 98 or 99% (basedon amount of sucrose). Most preferably, the obtainedtrehalulose-containing composition, that means vitalose, is essentiallyfree of sucrose, preferably is free of sucrose.

In a preferred embodiment of the present invention the cells, which maybe living or dead, or the cell extract may be used in non-immobilizedform. In a furthermore preferred embodiment the cells or the cellextracts, in particular the enzymes, are immobilized on at least onecarrier, preferably on two different types of carriers. In accordancewith this preferred embodiment one carrier immobilizes the cell or cellextract from the microorganism of the genus Pseudomonas and the otherdifferent carrier immobilizes the cells or cell extract from themicroorganism of the genus Protaminobacter.

In a particularly preferred embodiment the cells or cell extracts fromthe microorganisms of the genera Pseudomonas and Protaminobacter areco-immobilized on the same carrier. Thus, in this embodiment one and thesame carrier, i.e. preferably one type of carrier particles, do containboth cells or cell extracts combined on or in the carrier.

The immobilization can be carried out using prior art technologies.Preferably, the present invention foresees to use entrapment within agel. Cells can preferably also be physically adsorbed on an innersupport. They may preferably also be covalently coupled to an innersupport or they can be aggregated by use of a cross-linking agent.According to the present invention, entrapment in a gel is preferred.Suitable gel materials may be alginate, polyacrylamide, agar, xanthangum/locust bean gum, kappacarrageenan or kappacarrageenan/locust beangum, collagen or cellulose acetate.

In a preferred embodiment, the present invention foresees to useimmobilization of the cells or cell extracts in an alginate carrier, inparticular a calcium alginate. A preferred immobilization produces aninert three-dimensional polymer network, providing a high inwarddiffusion of sucrose and a high stability. To prepare in a preferredembodiment the immobilized cells or cell extracts, preferably in analginate gel, the cells or the cell extracts, preferably of both generaof microorganisms together, are mixed with an aqueous solution of asoluble alginate, for instance sodium alginate. In a preferred processthis will involve slurrying the whole cells or disrupted cells, i.e.cell extracts, with a soluble alginate. In a preferred embodiment, theconcentration of cells is between 1 and 90 weight-% of the solution,preferably 10 to 40 weight-%, preferably 20 weight-% (each net weight(volume)) of the solution. The resultant alginate mixture is thenmetered into a solution of a metal salt with which the soluble alginateforms a gel, for instance a calcium alginate produced by using calciumchloride. It is also possible to immobilize the microorganisms of thegenus Pseudomonas separately from the microorganisms of the genusProtaminobacter and mix the both separately immobilized microorganismsat a later stage before the sucrose conversion starts.

By metering the slurry or other alginate mixture as discrete droplets itis in a preferred embodiment possible to produce spherical pellets ofgel entrapping the cells or cell extracts. Preferably the pellet sizecan be varied, wherein pellets having a size of 3 to 5 diameter arepreferred. In a furthermore preferred embodiment it is also possible toimmobilize the cells or cell extracts in a block of gel, a rope of gelor in microfibrous particles. It is also possible to adsorb cells orcell extracts on DEAE-cellulose or by crosslink the cells or cellextracts, for instance with glutaraldehyde.

In a particularly preferred embodiment of the present invention it isforeseen to employ in the process of the present invention a specificratio, in particular ratio of enzymatic activities or weight ratio ofthe cells or cell extracts, of Pseudomonas and Protaminobacter. In aparticularly preferred embodiment a process for the production of atrehalulose-containing composition, that means vitalose, is providedwherein the weight ratio or enzymatic activity ratio of cells or cellextracts of Pseudomonas to Protaminobacter is from 10:1 to 1:10, inparticular from 8:1 to 1:8, from 5:1 to 1:5, from 3:1 to 1:3, from 2:1to 1:2 and in particular is 1:1. In a furthermore preferred embodimentof the present invention a process for the production oftrehalulose-containing composition, that means vitalose, is provided,wherein the ratio of weight or enzymatic activity of Pseudomonas toProtaminobacter is from 10:1 to 1.1:1, 8:1 to 1.1:1, 5:1 to 1.1:1, 3:1to 1.1:1, preferably from 2:1 to 1.1:1. In a furthermore preferredembodiment of the present invention a process for the production oftrehalulose-containing composition, that means vitalose, is provided,wherein the ratio of weight or enzymatic activity of Pseudomonas toProtaminobacter is from 1:1.1 to 1:10, 1:1.1 to 1:8, 1:1.1 to 1:5, 1:1.1to 1:3, 1:1.1 to 1:2. In a particularly preferred embodiment a processis provided according to the above, wherein the ratio of weight orenzymatic activities of cells or cell extracts from Pseudomonas toProtaminobacter is different from a 1:1 ratio, in particular, whereinthe ratio is greater than 1, that means >1:1 or is smaller than 1 thatmeans <1:1.

A process in accordance with the present invention is preferably carriedout as a continuous process, a discontinuous process, preferably inbatch wise operation or a semi-continuous process. The process maypreferably be carried out in a fixed bed reactor. The process maypreferably be carried out in a reaction vessel, a biofermentor or atank, preferably under mechanical agitation.

In one further embodiment the immobilized cells or cell extracts areloaded into a column and the substrate, i.e. the sucrose-containingsolution or suspension is passed through the column so as to harvest thetrehalulose-containing composition in the flow-through. Preferablyseveral columns may be employed in parallel, for example on a carrousel.

The present invention also provides a trehalulose-containingcomposition, that means vitalose, comprising trehalulose andisomaltulose, in particular obtained or being obtainable according toany one of the preceding processes. In particular, the combined andsimultaneous use of cells or cell extracts from the microorganism of thegenera Pseudomonas and Protaminobacter converts the sucrose-containingcomposition favourably in a specific trehalulose-containing composition,that means vitalose. Said composition, which is a natural, essentiallynon-cariogenic, low insulinemic and low glycemic sweetener composition,provides the physiologically advantageous properties as disclosedtherein and simultaneously provide a sucrose-like temporal and flavourprofile, which is not found in compositions comprising isomaltulose ortrehalulose each alone. Preferably, the trehalulose-containingcomposition, that means vitalose, of the present invention is used in aliquid form, in particular in form of a syrup, which advantageously in aparticularly preferred embodiment does not crystallize out. Trehaluloseand the trehalulose-containing composition, that means vitalose, of thepresent invention have a good solubility in aqueous media, which enablesa particularly wide range of applications.

In the context of the present invention a trehalulose-containingcomposition, that means vitalose, of the present invention is acomposition, which primarily comprises trehalulose and isomaltulose andwhich comprises minor components, which are in particular isomaltose,isomelezitose, carbohydrate oligomers with a DP (degree ofpolymerization) greater or equal to 3, and optionally glucose, fructoseand/or sucrose. In one embodiment of the present invention, it isforeseen to remove one, more or all of the minor components, forinstance isomaltose, isomelezitose and/or the oligomers. In oneembodiment, the sweetener composition of the present invention exhibitsa more sucrose-like temporal and/or sucrose-like flavor profile than asweetener composition comprising trehalulose or isomaltulose each alone.As used herein, the phrases “sucrose-like characteristic,” “sucrose-liketaste,” “sucrose-like sweet,” and “sucrose-like” are synonymous and arealways understood to relate to sucrose. Sucrose-like characteristicsinclude any characteristic similar to that of sucrose and include, butare not limited to, maximal response, flavor profile, temporal profile,adaptation behavior, mouthfeel, concentration/response functionbehavior, tastant and flavor/sweet taste interactions, spatial patternselectivity, and temperature effects. Whether or not a characteristic ismore sucrose-like, isomaltulose-like or trehalulose-like is determinedby expert sensory panel assessments of sucrose and the compositionscomprising trehalulose and/or isomaltulose. Such assessments quantifysimilarities of the characteristics of compositions comprisingtrehalulose and/or isomaltulose, with those comprising sucrose.

In a particularly preferred embodiment of the present invention thetrehalulose-containing composition, that means vitalose, preparedaccording to the present invention comprises a trehalulose contentsuitable to achieve the advantageous physiological properties describedherein, in particular the low insulin response induced in the consumer'sblood. In a particularly preferred embodiment the trehalulose-containingcomposition, that means vitalose, of the present invention comprisestrehalulose and isomaltulose each in a content that the composition issuitable to achieve the advantageous physiological properties describedherein, namely the low insulin response induced in the consumer's blood.In a particularly preferred embodiment of the present invention theproduct for human or animal consumption of the present inventioncomprises the trehalulose-containing composition, that means vitalose,of the present invention in an amount suitable to achieve theadvantageous physiological properties described herein in the consumer,in particular induces the desired low insulin response in the consumer'sblood.

In a particularly preferred embodiment the trehalulose-containingcomposition, that means vitalose, prepared according to the presentinvention comprises 20 to 95 weight-%, preferably 30 to 90 weight-%,preferably 25 to 75 weight-%, preferably 40 to 80 weight-% and inparticular 30 to 70 weight-%, preferably 35 to 65 weight-%, preferably45 to 70 weight-% trehalulose (based on dry matter of the composition).Most preferably, the trehalulose-containing composition, that meansvitalose, comprises 70 to 80 weight-%, preferably 65 to 78 weight-%trehalulose. Preferably, the remainder of the composition adds up to100% with isomaltulose, or, optionally isomaltulose and 0.1 to 1, 2, 3or 4 weight-% of the minor components.

In a furthermore preferred embodiment the trehalulose-containingcomposition, that means vitalose, of the present invention comprises 8,10, 20, or 35 to 50 weight-%, in particular 9 to 40 weight-%, preferably9 to 30 weight-%, preferably 22 to 35 weight-%, preferably 20 to 40weight-%, preferably 20 to 30 weight-%, preferably 18 to 27 weight-% andmost preferably 20 to 25 weight-% isomaltulose (based on dry matter ofthe composition). Preferably, the remainder of the composition adds upto 100% with trehalulose, or, optionally trehalulose and 0.1 to 1, 2, 3or 4 weight-% of the minor components.

In a furthermore preferred embodiment of the present invention thetrehalulose-containing composition, that means vitalose, of the presentinvention comprises 0.0, preferably 0.1 to 2.0, 0.1 to 1.5, preferably0.1 to 1.0, preferably 0.1 to 0.4 weight-% glucose.

In a furthermore, preferred embodiment of the present invention thetrehalulose-containing composition, that means vitalose, of the presentinvention comprises 0.0, preferably 0.1 to 2.0, 0.1 to 1.5, preferably0.1 to 1.0, preferably 0.1 to 0.4 weight-% fructose.

In a furthermore, preferred embodiment of the present invention thetrehalulose-containing composition, that means vitalose, of the presentinvention comprises 0.0, preferably 0.1 to 2.0, 0.1 to 1.5, preferably0.1 to 1.0, preferably 0.1 to 0.4 weight-% isomaltose.

In a furthermore, preferred embodiment of the present invention thetrehalulose-containing composition, that means vitalose, of the presentinvention comprises 0.0, preferably 0.1 to 2.0, 0.1 to 1.5, preferably0.1 to 1.0, preferably 0.1 to 0.4 weight-% isomelezitose.

In a furthermore, preferred embodiment of the present invention thetrehalulose-containing composition, that means vitalose, of the presentinvention comprises 0.0, preferably 0.1 to 2.0, 0.1 to 1.5, preferably0.1 to 1.0, preferably 0.1 to 0.4 weight-% carbohydrate oligomers,preferably with a DP≧3.

In a particularly preferred embodiment the trehalulose-containingcomposition, that means vitalose, prepared according to the presentinvention comprises 70 to 80 weight-% trehalulose and 20 to 30 weight-%isomaltulose. In a particularly preferred embodiment thetrehalulose-containing composition, that means vitalose, does notcomprise any other components than isomaltulose and trehalulose. Inanother preferred embodiment the trehalulose-containing composition,that means vitalose, prepared according to the present inventioncomprises 70 to 80 weight-% trehalulose and 20 to 30 weight-%isomaltulose and one or more of the carbohydrates selected from thegroup consisting of isomaltose, isomelezitose and carbohydrate oligomersadding up to 100 weight-% from the dry matter of the composition,preferably in a sum of 1 to 6, 1 to 5, 1 to 4 and most preferred 1 to 3weight-% (based on weight of composition). In a furthermore preferredembodiment the trehalulose-containing composition, that means vitalose,comprises also isomaltose, isomelezitose, glucose, fructose, sucrose andcarbohydrate oligomers, preferably in an overall sum adding up to 100weight-% (DM), preferably in a sum of 1 to 6, 1 to 5, 1 to 4 and mostpreferred 1 to 3 weight-% (based on weight of composition).

In a preferred embodiment of the present invention it is foreseen toremove one or more of the minor components of the obtainedtrehalulose-containing composition, that means vitalose, preferably oneor more of the group consisting of sucrose, glucose, fructose,isomaltose, isomelezitose and carbohydrate oligomers, most preferablyfrom the group consisting of glucose, fructose and sucrose. In apreferred embodiment said removal may be carried out by methods known inthe art, such as filtration, chromatography, for instance anion and/orcation exchange chromatography and enzymatic, for instance invertase, orcatalytic cleavage. Preferably, a charcoal and/or resin treatment may becarried out. Thus, in a particularly preferred embodiment it is possibleto produce a trehalulose-containing composition, that means vitalose,being free of glucose, fructose and sucrose, which is completelynon-cariogenic and provides a particularly low glycemic and insulinemicindex.

In a furthermore preferred embodiment of the present invention there isprovided a product for human or animal consumption, comprising thetrehalulose-containing composition, that means vitalose, of the presentinvention and at least one additive.

In a preferred embodiment of the present invention the product for humanor animal consumption comprises 0.02 to 3.0 weight-%, in particular 0.02to 1.0 weight-%, preferably 0.02 to 0.5 weight-% of the at least oneadditive and an amount of the trehalulose-containing composition, thatmeans vitalose, adding up to 100%, in particular 97 to 99.98 weight-%,preferably 99.0 to 99.98 weight-%, in particular 99.5 to 99.98 weight-%of the trehalulose-containing composition, that means vitalose (eachbased on dry matter). Preferably, the at least one additive contained inthe product, preferably in an amount from 0.02 to 3.0 weight-%, is (a) astevia extract or a steviolglycoside, (b) a stevia extract or asteviolglycoside and at least one of lactobionic acid,lactobionic-δ-lactone, a salt of lactobionic acid or a mixture thereof,or (c) at least one of a lactobionic acid, lactobionic-δ-lactone, a saltof lactobionic acid or a mixture thereof.

In a preferred embodiment of the present invention the product for humanor animal consumption comprises 3 to 95 weight-%, preferably 5 to 95weight-%, preferably 20 to 95 weight-%, preferably 5 to 90 weight-%,preferably 10 to 90 weight-%, preferably 20 to 80 weight-%, preferably 7to 70 weight-%, most particularly 40 to 60 weight-% of thetrehalulose-containing composition, that means vitalose, of the presentinvention and 5 to 97 weight-%, preferably 5 to 95 weight-%, mostpreferably 5 to 80 weight-%, preferably the remainder adding up to 100weight-%, of the at least one additive (each based on dry matter on theoverall product).

In a preferred embodiment of the present invention the additive is ahigh intensity sweetener, preferably is at least one of a stevia extractor a steviolglycoside, in particular a rebaudioside, preferablyrebaudioside A (Reb A, CAS no. 58 543-16-1, C₄₄H₇₀O₂₃). Most preferred,the trehalulose-containing composition, that means vitalose, of thepresent invention is present in a product for human or animalconsumption together with Reb A as an additive.

For example, particular embodiments may also comprise combinations ofsteviolglycosides.

Non-limiting examples of suitable steviolglycosides which may or may notbe combined include rebaudioside A, rebaudioside B, rebaudioside C,rebaudioside D, rebaudioside E, rebaudioside F, dulcoside A, dulcosideB, rubusoside, stevioside (CAS no. 57817-89-7, C₃₈H₆₀O₁₈), orsteviolbioside. According to particularly desirable embodiments of thepresent invention, the combination of high intensity sweetenerscomprises rebaudioside A in combination with rebaudioside B,rebaudioside C, rebaudioside E, rebaudioside F, stevioside,steviolbioside, dulcoside A or combinations thereof. For the chemicalnomenclature it is referred to the 68^(th) JECFA meeting (2007)published in FAO JECFA Monographs 4 (2007).

In a particularly preferred embodiment a steviolglycoside used as a highintensity sweetener additive according to the present invention is atleast one of the group consisting of stevioside, rebaudioside A,rebaudioside C, dulcoside A, rubusoside, steviolbioside and rebaudiosideB.

In a particularly preferred embodiment of the present invention the highintensity sweetener used in the present invention is a mixture of Reb A,rebaudioside B, rebaudioside C, dulcoside A, rubusoside, steviolbiosideand stevioside, which comprises at least 95 weight-% (based on drymatter of the sweetener composition) of said steviolglycosides.

In a particularly preferred embodiment of the present invention a highintensity sweetener comprising rebaudioside A is used, wherein therebaudioside A content is at least 97 weight-%, preferably is >97weight-% of the dry weight of the sweetener composition.

According to a particularly desirable embodiment, the high intensitysweetener comprises a mixture of rebaudioside A, stevioside,rebaudioside B, rebaudioside C, and rebaudioside F; wherein rebaudiosideA is present in the mixture of high intensity sweeteners in an amount inthe range of about 75 to about 85 weight percent based on the totalweight of the mixture of high intensity sweeteners, stevioside ispresent in an amount in the range of about 1 to about 6 weight percent,rebaudioside B is present in an amount in the range of about 2 to about5 weight percent, rebaudioside C is present in an amount in the range ofabout 3 to about 8 weight percent, and rebaudioside F is present in anamount in the range of about 0.1 to about 2 weight percent.

According to particularly preferred embodiments, the purity of therebaudioside A used may range from about 50% to about 100%; from about70% to about 100%; from about 80% to about 100%; from about 90% to about100%; from about 95% to about 100%; from about 95% to about 99.5%; about96% to about 100%; from about 97% to about 100%; from about 98% to about100%; and from about 99% to about 100%. According to particularlypreferred embodiments, upon crystallization of crude rebaudioside A, thesubstantially pure rebaudioside A composition comprises rebaudioside Ain a purity greater than about 95% by weight up to about 100% by weighton a dry basis. In other preferred embodiments, substantially purerebaudioside A comprises purity levels of rebaudioside A greater thanabout 97% up to about 100% rebaudioside A by weight on a dry basis,greater than about 98% up to about 100% by weight on a dry basis, orgreater than about 99% up to about 100% by weight on a dry basis.

In a furthermore preferred embodiment of the present invention theadditive is at least one of lactobionic acid, lactobionic-δ-lacton, asalt of lactobionic acid or a mixture thereof. According to the presentinvention the use of these lactobionic acid-based additives provides theadvantage of masking or reducing the bitter off-taste of otherwise sweetcompounds and provides a sweet flavouring enhancing effect.

Lactobionic acid, lactobionic-δ-lactone, a salt of lactobionic acid ormixtures thereof are thus capable of masking for example bitterflavours. One preferred embodiment of the present invention is a maskingof bitter flavours, in particular attributable to stevia extracts or asteviolglycoside, for example rebaudioside A, wherein lactobionic acid,lactobionic-δ-lactone, a salt of lactobionic acid or mixtures thereofare used in combination with the trehalulose-containing composition,that means vitalose, of the present invention and stevia extracts or asteviolglycoside, for instance Reb A, of which it is known that theyshow a bitter off-taste.

Thus, preferably, the trehalulose-containing composition, that meansvitalose, is combined with a stevia extract or a steviolglycoside,particularly a rebaudioside, in particular rebaudioside A, and withanother additive, in particular at least one of lactobionic acid,lactobionic-δ-lacton, a salt of lactobionic acid or a mixture thereof.

In a preferred embodiment the product for human or animal consumptiontherefore comprises the trehalulose-containing composition, that meansvitalose, of the present invention and at least two additives, namely astevia extract or a steviolglycoside, in particular Reb A, and at leastone of lactobionic acid, lactobionic-δ-lactone or a salt or mixturethereof. Such a product displays a similar sweetening power thanconventional sucrose or sucrose/glucose-based products, but has thephysiologically advantageous properties disclosed herein and does notshow any unfavourable organoleptic or taste features known to beattributable to conventional artificial sweeteners. In particular, sucha product provides a high nutritional and even therapeutical value incombination with a sucrose-like profile of characteristics, inparticular sucrose-like taste and sweetness.

The amount of lactobionic acid, lactobionic-δ-lactone, a salt oflactobionic or mixtures thereof that may preferably be incorporated intoa food composition in order to benefit from its flavour-enhancing and/ormasking properties may vary within wide ranges, and lies preferablybetween 0.01 wt. % and 90 wt. %, based on the whole of the foodcomposition. Preferably, the amount of lactobionic acid,lactobionic-δ-lactone, a salt of lactobionic acid or mixture thereofthat is incorporated into a food composition is at least 0.05, 0.10,0.25, 0.50, or 1.00 wt. %, based on the whole of the food composition.Preferably the amount of lactobionic acid, lactobionic-δ-lactone, a saltof lactobionic acid or mixture thereof that is incorporated into a foodcomposition is at most 95, 85, 75, 65, 55, 45, 40, 35, 30, 25, 20, 15,or 10 wt. %, based on the whole of the food composition. Dosages oflactobionic acid, lactobionic-δ-lactone, a salt of lactobionic acid ormixtures thereof in amounts of 5 or 10 wt. % to 90 wt. % are oftensuitable for use in concentrates, table top sweeteners, and saltreplacements. Moreover, it was found that the said high dosages oflactobionic acid, lactobionic-δ-lactone, a salt of lactobionic acid ormixtures thereof can be associated with its use as a carrier, forexample a carrier of aromas.

If lactobionic acid is used as such, then it may lead to the formationof lactobionic-δ-lactone. In that case, the ratio between lactobionicacid and lactobionic-δ-lactone is determined at least partly by thespecific conditions of use. By analogy, the use of lactobionic-δ-lactoneas such may lead to the formation of lactobionic acid. The saidformation and subsequent presence of lactobionic-δ-lactone orlactobionic acid are understood to be according the use of the presentinvention.

If a mixture of lactobionic acid, lactobionic-δ-lactone and a salt oflactobionic acid is used in a preferred embodiment then the weight ratioof, one the one hand, the sum of lactobionic acid andlactobionic-δ-lactone to, on the other hand, the salt of lactobionicacid in the mixture, lies preferably between 1:10 and 10:1.

In a furthermore preferred embodiment of the present invention theadditive is selected from the group consisting of acidic flavours, fruitflavours, sweet flavours, savoury flavours, salty flavours, a highintensity sweetener, a sugar alcohol, sucromalt, ribose, tagatose,trehalose, organic acid, e.g. citric acid or lactic acid, fruitextracts, a bulk sweetener, a fibre, a prebiotic agent, a thickener, avitamin, a mineral, a preservative, a fruit preparation, a food colour,a liquid medium, such as water or milk, and a therapeutic agent.

In a furthermore preferred embodiment of the present invention theadditive is selected from the group consisting of creatine, polyphenol,L-carnitin, omega-3 polyunsaturated fatty acid, omega-6 polyunsaturatedfatty acid, green tea extract, EGCG (epigallocatechingallate),aminoacids and peptopro, which is a pre-digested casein-derived milkprotein.

In a further particularly preferred embodiment as a high intensitysweetener aspartame, monellin, brazzeine, cyclamate, neotame, acesulfameK, glycyrrhicine, saccharine, sucralose, alitame,neohesperidine-dihydrochalcone, stevioside, Reb A or thaumatin can beused.

Thus, the trehalulose-containing composition, that means vitalose, mayalso be combined with an additive, which is at least one sugar alcohol,preferably selected from the group consisting of isomalt, isomalt ST,isomalt GS, mannitol, sorbitol, xylitol, lactitol, erythritol ormaltitol.

In a preferred embodiment, as an additive a fibre, preferably a solublefibre, particularly resistant dextrines, resistant maltodextrines, forinstance nutriose, pectine, carrageenan, rice starch, rice syrup,polydextrose, oligosaccharides, for instance galactooligosaccharides orfructooligosaccharides, or fructans, for instance inulin, can be used.

In a furthermore preferred embodiment of the present invention theadditive is a tryptophane-containing peptide composition, in particulartryptophane-containing peptides or tryptophane itself. In a preferredembodiment the tryptophane-containing peptide composition comprises atleast two different tryptophane-containing peptides, preferablywater-soluble peptides. In a particularly preferred embodiment thetryptophane-containing peptides are di- or tripeptides. In aparticularly preferred embodiment a food, feed or pharmaceutical productcomprising vitalose and the tryptophane-containing peptide compositioncomprises 0.5 to 5 weight %, preferably 0.9 to 3.5 weight % (dry weight)of the tryptophane-containing peptide composition. In a particularlypreferred embodiment of the present invention such a product isparticularly useful for improving the mental performance of a subject.

The present invention also provides a product for human and animalconsumption which is a food, an animal feed or a pharmaceutical product.The product according to the present invention is preferably aconfectionery, a filling for a confectionery, a soft caramel, a hardcaramel, a fondant, a yoghurt, a bakery product, a chewing gum, an icecream, a milk product, a jelly, a beverage, a fruit juice concentrate, afruit preparation, a marmalade or a smoothie. Preferably, the product isa fruit preparation, a fruit juice, a jelly or a fondant. The beveragemay preferably be a beer, a fruit juice, a milk drink, a soft drink, anisotonic drink, a hypertonic drink, a cacao drink, a rice drink, a soyadrink, an alcohol free beverage, a beverage with heat- and pH-stableosmolarity, an energy drink or a sport drink.

In a preferred embodiment of the present invention the product for humanor animal consumption is a beverage, in particular a beverage comprisingat least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60 or 70weight-% of the trehalulose-containing composition, that means vitalose,optionally including at least one additive, and at most 10, 20, 30, 40,50, 60, 70, 75 or 80 weight-% (each based on total weight of thebeverage) of the trehalulose-containing composition, that meansvitalose, optionally including at least one additive, of the presentinvention the remainder being a liquid medium, in particular an aqueousor milky solution, such as water or milk. In a particularly preferredembodiment the beverage comprises 1 to 30%, preferably 2 to 25%, mostpreferably 3 to 15% (each based on total weight of the beverage) of thetrehalulose containing composition, that means vitalose, optionallyincluding at least one additive, of the present invention the remainderbeing a liquid medium, in particular an aqueous or milky solution, suchas water or milk. Thus, in a preferred embodiment the beverage maycontain one or more of the above identified additives, in particular (a)a stevia extract or a steviolglycoside, preferably Reb A, or (b) alactobionic acid, lactobionic-δ-lacton, a salt of lactobionic acid or amixture thereof or both of them, that means (a) and (b). In aparticularly preferred embodiment of the present invention thetrehalulose-containing composition, that means vitalose, of the presentinvention or a product comprising the trehalulose-containingcomposition, that means vitalose, of the present invention does notcomprise any further sweetening agent except the trehalulose-containingcomposition, that means vitalose, itself. In a furthermore preferredembodiment of the present invention the trehalulose-containingcomposition, that means vitalose, of the present invention or a productcontaining said composition does not comprise any other sugar besidesthe sugars contained in the trehalulose-containing composition thatmeans vitalose.

In a furthermore preferred embodiment of the present invention thetrehalulose-containing composition, that means vitalose, of the presentinvention or a product containing the trehalulose-containingcomposition, that means vitalose, of the present invention is free ofhigh intensity sweeteners, is free of sugar replacement agents, such assugar alcohols, or is free of both. In a particularly preferredembodiment of the present invention the trehalulose-containingcomposition, that means vitalose, or a product containing saidcomposition is free of sorbitol, is free of mannitol, is free oflactitol, is free of maltitol, is free of xylitol, is free of erythritolor is free of two, three, four, five or all of them.

The present invention also relates to the use of thetrehalulose-containing composition, that means vitalose, of the presentinvention or trehalulose itself for preparing a product for human oranimal consumption, which product is capable and designed for asustained release of glucose and for simultaneously eliciting (hereinalso called inducing) in a consumer a low blood insulin response, inparticular eliciting an insulin response which is significantly lowerthan elicited by isomaltulose alone.

The present invention also relates to the above-mentioned use, whereinthe product is a pharmaceutical product.

The present invention relates to a method for treating a glucose orinsulin metabolism-related condition or disease, wherein the product isapplied to a subject in need thereof, thereby eliciting a low insulinresponse and a sustained release of glucose.

The present invention relates to the above method, wherein the glucoseor insulin related condition or disease is selected from the groupconsisting of diabetes, metabolism syndrome, glucose intolerance,insulin resistance, adipositas, obesity, hyperlipidemia, cancer, inparticular colon cancer, a liver disease, insulin sensitivity andarterioscleroses.

The present invention is thus also based upon the unexpected findingthat trehalulose provides upon uptake by a human or animal consumer incomparison to isomaltulose an increased level of blood glucose andsimultaneously elicits a, in comparison to isomaltulose, lower level ofinsulin in the blood. Thus, although isomaltulose and trehalulose bothare low glycemic sugars, trehalulose unexpectedly and advantageouslyprovides the advantage of providing in contrast to isomaltulose a higherlevel of blood glucose upon ingestion in the human or animal body andsimultaneously elicits a lower insulin response, i.e. a lower insulinlevel in the blood, in contrast to isomaltulose. Thus, trehalulose is aparticularly low glycemic agent with a low GI (glycemic index) providingan extremely low insulin response, i.e. provide a very low II (insulinindex).

In the context of the present invention a “low insulin response” is aninsulin response, which is induced by trehalulose and which is lowerthan the insulin response induced by isomaltulose. The present inventiontherefore provides the unexpected teaching that isomaltulose andtrehalulose both have a low GI and that trehalulose has a particularlylow insulin index (II), meaning provides a low insulin response, i.e. aninsulin response lower than that of isomaltulose. The present inventiontherefore opens up new and advantageous methods and applications fortrehalulose and trehalulose-containing compositions, which make use ofthe above-identified finding. In particular the present inventionprovides the teaching to use trehalulose or a trehalulose containingcomposition, that means vitalose, for the preparation of products, inparticular for human and animal consumption, which products are designedand suitable for providing a sustained release of glucose andsimultaneously elicit only low insulin response, in contrast to a foodcomprising isomaltulose alone. Thus, a food according to the presentinvention aims to reduce the level of insulin in the blood and helps toprevent and treat obesity, overweight, adipositas, diabetes mellitus andother glucose and insulin-related diseases or conditions.

Thus, a composition or a food according to the present inventioncomprising the identified amounts of trehalulose is unexpectedlyadvantageous in comparison to known compositions, in particular solelyor primarily isomaltulose-containing compositions or products, interalia due to its low insulinemic index. Thus, the present inventionprovides the technical teaching so as to specifically use and adapt atrehalulose content in a composition, food or method of treatment so asto replace other sweeteners, in particular sucrose, glucose and/orisomaltulose, completely or partially so as to provide a foodcomposition or therapy supplying comparable high amount of glucose andthereby providing a very low insulin response in the consumer's blood.Thus, the present invention provides the advantageous teaching that fora certain level of energy supply a comparable lower insulinemic load isburdened onto the consumer or vice versa that with a certain insulinemicload a higher amount of energy can be supplied to the consumer by usingthe specific teaching of the present invention, that means theparticular properties of trehalulose to provide an extremely lowglycemic index and a comparable higher glucose supply to the consumer incontrast to isomaltulose.

The present invention therefore also provides products, uses and methodsfor treating glucose or insulin metabolism-related conditions ordiseases, wherein a product according to the present invention isapplied in an amount suitable to achieve the desired effect to a subjectin need thereof, in particular a human or animal being thereby elicitinga low insulin response and a sustained release of glucose. In thecontext of the present invention a glucose or insulin metabolism-relateddisease is selected from the group consisting of diabetes, in particulardiabetes mellitus, type II, metabolism syndrome, glucose intolerance,insulin resistance, insulin sensitivity, adipositas, obesity,hyperlipidemia, cancer, liver diseases, colon cancer and overweight. Thepresent invention provides also the teaching that the use of trehaluloseor trehalulose-containing composition, that means vitalose, of thepresent invention increases the physical and mental performance,increases fat consumption and oxidation, controls the blood glucose andinsulin levels, mobilizes free fatty acids and reduces the consumptionof glycogen.

The present invention provides the advantageous teaching thattrehalulose or the trehalulose-containing compositions, that meansvitalose, of the present invention are capable of continuously supplyingthe body of the consumer, in particular the brain, nerve, blood andmuscle cells with glucose for a prolonged and sustained time in a leveleven higher than provided by isomaltulose while simultaneously, inparticular in comparison to isomaltulose, a comparable low insulinresponse is induced. Thus, the body is provided with a long lasting andcontinuous energy supply avoiding a reduction in the physical and mentalperformance and increasing the physical and mental endurance, conditionand activity, while reducing the consumption of the glycogen reservesand supporting weight control and weight reduction.

The present invention also relates to a method for the prophylaxes andtreatment of tooth diseases, in particular plaque formation or toothdefects, in particular caries, wherein a product or composition of thepresent invention is applied in an amount suitable to achieve thedesired effect to a subject in need thereof. In particular, the presentinvention, according to which a product or a composition of the presentinvention is applied to a subject in need thereof, reduces the plaqueformation in the mouth of the consumer. Thus, the present inventionprovides a method for improving the tooth health.

In a furthermore preferred embodiment the present invention relates to amethod for improving the mental and/or physical performance, inparticular the endurance, wherein a product or composition according tothe present invention is applied in an amount suitable to achieve thedesired effect to a subject in need thereof. Thus, the present inventionprovides in a particularly preferred embodiment a method for improvingthe endurance of a subject in need thereof, whereby in a particularlypreferred embodiment the glycogen consumption is reduced.

In a furthermore preferred embodiment the present invention relates to amethod for improving, preferably increasing, the fat oxidation of asubject in need thereof, whereby a product or composition according tothe present invention is applied in an amount suitable to achieve thedesired effect to a subject in need thereof. Furthermore, the presentinvention relates to a method for the management of body weight, whereina composition or a product according to the present invention is appliedto a subject in need thereof.

In a particularly preferred embodiment the present invention provides amethod for reducing the respiratory quotient (RQ=CO₂ eliminated/O₂consumed) of a patient, wherein the product or a composition accordingto the present invention is applied in an amount suitable to achieve thedesired effect prior, during or after a physical exercise to the subjectin need thereof.

All the methods of using the products and compositions of the presentinvention can be therapeutic or non-therapeutic methods, for instancerelating to cosmetic, activity-related, athletic or life-styleorientated methods.

Further preferred embodiments are the subject matter of the subclaims.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1 to 6 show a graphical presentation of the conversion of asucrose-containing composition into a trehalulose-containingcomposition, that means vitalose, of the present invention for variousratios of Pseudomonas mesacidophila and Protaminobacter rubrum,

FIGS. 7 to 14 show graphically the hardness and cohesiveness of jelliesprepared according to the present invention in comparison toconventional sucrose-based jellies,

FIG. 15 shows the blood glucose level of a consumer after consumption ofisomaltulose and trehalulose and

FIG. 16 shows the blood insulin level of a consumer after consumption ofisomaltulose and trehalulose.

EXAMPLES

The following examples are provided only for the purpose of illustratingthe invention. They are not to be construed in any way as limiting.

Example 1 Preparation of Vitalose A. Preparation of the ProtaminobacterBiocatalyst

Cells are rinsed off a subculture of the strain Protaminobacter rubrum(CBS 574.77) with 10 ml of a sterile nutrient substrate composed of 8 kgof thick juice from a sugar works (dry matter content 65%), 2 kg of cornsteep liquor, 0.1 kg of (NH₄)₂HPO₄ and 89.9 kg of distilled water,adjusted to pH 7.2 if required. This suspension is used as inoculum forthe shaker preculture in 1 liter flasks containing 200 ml of nutrientsolution of the above composition.

After an incubation time of 30 hours at 29° C., 18 liters of nutrientsolution of the above composition in a 30 liter small fermenter areinoculated with, in each case, 10 flasks (total content 2 liters) andcultivated at 29° C. with 20 liters of air per minute and a stirrerspeed of 350 rpm.

After the organism counts have reached above 5×10⁹ organisms/ml, thecultivation is stopped, the cells are harvested from the fermentersolution by centrifugation, suspended in a 2% strength sodium alginatesolution and immobilized by adding the suspension dropwise to a 2%strength calcium chloride solution.

The resulting immobilisate beads are washed with water. This biocatalystcan be stored at +4° C. for several weeks.

B. Preparation of the Pseudomonas Biocatalyst

To prepare this biocatalyst, cells were rinsed off a subculture of thestrain Pseudomonas mesoacidophila MX-45 (FERM BP 3619) with 10 ml of asterile nutrient substrate composed of 8 kg of thick juice from a sugarworks (dry matter content=65%), 2 kg of corn steep liquor, 0.1 kg of(NH₄)₂HP0₄ and 89.9 kg of distilled water, adjusted to pH 7.2 ifrequired. This suspension was used as inoculum for a shaker preculturein a 1 liter flask containing 200 ml of the nutrient solution.

After incubation at 29° C. for 30 hours, 18 liters of nutrient solutionof the above composition in a 30 liter small fermenter were inoculatedwith, in each case, 10 flasks (total content 2 liters) and cultivated at29° C. with 20 liters of air per minute and a stirrer speed of 350 rpm.

After the organism counts had reached above 5×10⁹ organisms/ml, thecultivation was stopped, the cells were harvested from the fermentersolution by centrifugation, suspended in a 2% strength sodium alginatesolution and immobilized by adding the suspension dropwise to a 2%strength calcium chloride solution. The resulting immobilisate beadswere washed with water. This biocatalyst can be stored at +4° C. forseveral weeks.

C. Preparation of Vitalose

The immobilized cells obtained as in A) are mixed in a 1:1 weight ratiowith the immobilized cells obtained as in B) and are together packed ina column reactor which can be thermostatted and are thermostatted at 25to 30° C., and a sucrose solution with a DM (dry matter) content of 35to 45% is passed through continuously. The flow rate in this case isadjusted so that at least 97% of the sucrose employed is converted intothe desired trehalulose-containing composition, that means vitalose.

HPLC analysis of the trehalulose-containing composition emerging fromthe column reactor revealed the following composition:

Fructose  1.0% of DM Glucose  1.0% of DM Sucrose  1.0% of DMIsomaltulose 22.9% of DM Trehalulose 73.3% of DM Isomaltose  0.5% of DMIsomelezitose  0.1% of DM Oligomers (DP > 3)  0.2% of DM.

D. Optional Removal of Remaining Sucrose

The remaining sucrose, was, optionally, removed from thetrehalulose-containing composition, that means vitalose, obtained inthis way by treating it in a strongly acidic cation exchanger loadedwith H⁺ ions or with suitable enzymes in a column reactor as follows:

i) Removal of the remaining sucrose on strongly acidic cation exchangers

100 cm³ of a strongly acidic cation exchanger (for example Lewatit® OC1052) were packed in a suitable glass column thermostatted at 60° C. andloaded with H⁺ ions by regeneration with HCl by a known method.

The trehalulose-containing composition, that means vitalose, obtained inExample 1C was pumped at a flow rate of 100 cm³·h⁻¹ through the cationexchanger column prepared in this way. The product obtained at thecolumn outlet had the following composition (HPLC):

Fructose  1.5% of DM Glucose  1.5% of DM Sucrose  0.0% of DMIsomaltulose 22.9% of DM Trehalulose 73.3% of DM Isomaltose  0.5% of DMIsomelezitose  0.1% of DM Oligomers (DP > 3)  0.2% of DMii) Alternatively, removal of the remaining sucrose was carried outusing an immobilized invertase (for example SP 362 from NOVO NORDISK A/SCopenhagen). 11 g of said immobilized enzyme corresponding to a bedvolume of 33 cm³ was packed in a suitable glass column thermostatted at60° C.

The trehalulose-containing composition, that means vitalose, obtained inExample 1 C was pumped at a flow rate of 210 cm³·h⁻¹ continuouslythrough this column.

HPLC analysis of the product emerging from the “invertase column”revealed a composition as indicated in section 1D i), above.

In both cases, the remaining sucrose was completely cleaved to glucoseand fructose. The content of these monosaccharides was correspondinglyhigher whereas the other components of the trehalulose-containingcomposition, that means vitalose, were unchanged.

It is also possible to remove glucose and fructose as well, so as toobtain a completely non-cariogenic sweetener.

Example 2 Sucrose Conversion with Various Ratios of Pseudomonas andProtaminobacter

In this example, the influence of various ratios of Pseudomonasmesoacidophila MX-45 (FERM BP 3619) and Protaminobacter rubrum CBS574.77 on the product composition was analysed.

To prepare the sucrose-containing composition in form of an aqueousmedium first solution A was prepared by preparing an aqueous solution of1.58 g/l calcium acetate (adjusted to a pH of 5.5 with an acetic acid).Then, solution B was prepared by adding solution A up to 1 l to 1.4602mol/l sucrose corresponding to 500 g sucrose.

Both strains of the microorganism, that means P. mesacidophila MX-45 andP. rubrum CBS 574.77, were cultivated separately as identified inexample 1 A and 1 B. After the organism counts each had reached above5×10⁹ organism/ml, the cultivation of both microorganisms was stopped,the cells were harvested by centrifugation and pooled together in a 2%strength sodium alginate solution and immobilized therefore together byadding the suspension drop wise to a 2% strength calcium chloridesolution. The weight ratio employed for combining the both strains ofmicroorganism on the alginate beads prepared thereby is apparent fromthe below, namely ratios of Pseudomonas to Protaminobacter of 1:2, 1:1and 2:1.

5 g of the biocatalyst prepared as indicated was mixed with 50 ml ofsolution A and kept at room temperature for three hours.

The conversion of the sucrose was started by mixing the suspension ofthe biocatalyst prepared above in a stirred reactor with 200 ml of thesucrose-containing solution B prepared above at 20° C.

In first set of experiments a biocatalyst was used, wherein from theoverall load of microorganism on the carriers 33% was P. mesoacidophilaMX-45 and 67% P. rubrum (% based on weight of the overall microorganismload). In a second set of experiments 50% of the overall microorganismload was MX-45 and 50% P. rubrum (% based on weight of the overallmicroorganism load). In a third set of experiments 67% of the overallload of microorganism was MX-45 and 33% P. rubrum (based on weight ofthe overall microorganism load).

Samples were taken after 5, 10, 15, 20, 25, 30, 35 and 60 minutes, 3hours and 20.5 hours. The samples taken were cooked, filtrated, dilutedto a concentration of 10° Bx (brix, 10 g in 100 g solution) and analysedby HPLC.

The results are given in FIGS. 1 to 6.

From FIGS. 1 to 6 it can be seen that the process of the presentinvention leads to a complete or almost complete conversion of thesucrose into a trehalulose-containing composition, that means vitalose.

In particular it can be seen that depending up on the specific weightratio between the cells of the two different microorganism strainsimmobilized varying ratios of trehalulose to isomaltulose can beobtained. In particular, the more P. mesoacidophila is used, the higherthe trehalulose content in the product composition is, while the higherthe P. rubrum proportion, the higher the isomaltulose content in theobtained product is. In particular, the data show that it is possibleand highly advantageous to use both microorganism and both enzymaticnutritives simultaneously together for the conversion and that nonegative effect on the overall conversion occurred.

Example 3 Preparation of Various Jellies

In the following, four recipes for jellies using as sweeteners a mixtureof sucrose and glucose and four recipes using the trehalulose-containingcomposition, that means vitalose, of the present invention are prepared.The jellies prepared were tested comparatively in storage andTPA-analysis.

3A: Recipes

Sample No. 1 2 3 4 Sucrose/ Sucrose/ Sucrose/ Sucrose/ 5 6 7 8 GlucoseGlucose Glucose Glucose Trehalulose Trehalulose Trehalulose Trehalulose(240 g dry (325 g dry (325 g dry (210 g dry (240 g dry (325 g dry (325 gdry (250 g dry substance, substance substance, substance, substance,substance, substance, substance, 41.5%) 56%) 60%) 24%) 41.5%) 56%) 60%)24%) Pectin H&F,  13.2 g  13.2 g AF 605 Citric acid,  5.0 g  6.0 g  5.0g  7.0 g anhydrous Water 290.0 g 104.8 g 128.8 g  39.3 g 232.5 g 27.0 g50.9 g Sugar 120.0 g 162.6 g 162.6 g 105.0 g Glucose 150.0 g 203.2 g203.2 g 131.3 g syrup DE 42 (approx. 80% dry substance) Trehalulose327.5 g 443.5 g  443.7 g  (73.3% dry substance) Trehalulose 344.7 g(73.1% dry substance) Lemon  0.4 g  0.4 g  0.4 g  0.5 g  0.4 g  0.4 g 0.4 g  0.6 g aroma Gelatine  35.0 g  18.0 g 35.0 g 18.0 g (280 Bloom)Water for  60.0 g 60.0 g gelatine Citric acid  10.0 g  10.0 g 10.0 g10.0 g solution, 50% Starch purity  18.0 g 18.0 g gum 400 Gum arabic600.0 g 720.0 g solution (52.9%) legend: Sample no. 1 to 4:sucrose/glucose compositions Sample no. 5 to 8: vitalose(trehalulose-containing composition) of the present invention

Composition of Sample 5 to 8:

water content (Karl Fischer- method) fructose glucose sucroseisomaltulose trehalulose isomaltose DP-3 isomelezitose remainder Sampleg/100 g No. titrim. HPLC-NH2 5 TREHALULOSE/ 44.5 0.1 0.2 <0.1 4.6 41.20.1 <0.1 <0.1 0.2 PECTINE 6 TREHALULOSE/ 21.2 0.2 0.3 <0.1 7.3 63.9 0.2<0.1 <0.1 0.4 GELATINE 7 TREHALULOSE/ 17.1 0.2 0.2 <0.1 7.5 67.4 0.2<0.1 <0.1 0.4 GELATINE/ STARCH 8 TREHALULOSE/ 22.7 0.3 0.3 <0.1 2.5 16.2<0.1 <0.1 <0.1 3 GUM ARABIC

3A: Recipes 3 B. TPA-Analysis

In a comparative testing the samples prepared in example 3A, above, wereanalysed in detail, i.e. the sucrose/glucose compositions containingvarious thickeners (pectin, gelatine, starch and/or gum arabic) werecompared to the trehalulose-containing compositions, that meansvitalose.

i) TPA-analysis (hardness, cohesiveness)ii) storage in closed PE(polyethylene)-bags at 25° C. for 8 weeks

The intervals of analysis for the TPA tests and the storage behaviourwere done after O (begin of storage), 1, 2, 4, 8 weeks.

In all of the jelly recipes the conventional sweetener combination ofsucrose and glucose could be successfully replaced by thetrehalulose-containing composition, that means vitalose, of the presentinvention. The preparation using the trehalulose-containing composition,that means vitalose, was exactly as for the sucrose/glucosecompositions. During the preparation, in particular when cooking thejellies, no degradation of trehalulose was observed.

As is evident from the results given in the FIGS. 7 to 14, the TPAmeasurements of hardness and cohesiveness show that the jellies madefrom the trehalulose-containing composition, that means vitalose, of thepresent invention are equal or very similar to the conventionalsucrose/glucose based products. The storage behaviour was alsocomparable. Storage at refrigerator temperatures, that means 7° C.,showed a particular good stability and prolonged shelf life.

Thus, these results show that both in terms of preparation and productcharacteristics the present trehalulose-containing composition, thatmeans vitalose, can be used instead of the conventionally usedsucrose/glucose sweeteners in jellies, but in addition provide theadvantages mentioned herein, namely the specific physiological andnutritional advantages in regard of the glycemic and insulinemicbehaviour.

Example 4 Preparation of Various Products with theTrehalulose-Containing Composition, that Means Vitalose, of the PresentInvention 4.1) Cereal Bar

Trehalulose - Raftilose ® bars Amount [g]   12% Trehalulose syrup 60.00(73.1% dry substance, see example 3) “trehalulose”* 4.42% water 22.0816.58%  Water (desalted) 82.92   21% Oligofructose P95 (Orafti)(Raftilose ®) 105.00   8% Fat Toffix P (Sasol) 40.00  1.9% EmulgentDimodan HP (Danisco) 9.50   10% Lactoprotein, Promilk 852 A1 (Ingredia)50.00 0.05% Ascorbic acid powder Reinst (Fluka) 0.25 0.05% Vitaminemixture (Döhler) 0.25   10% Desiccated coconut, Backfee 50.00   8%Chopped almonds, Backfee 40.00   8% Whole grain oat meal (Kölln) 40.00 100% In total 500.00 *Here “Trehalulose” refers to a 90:10trehalulose/isomaltulose mixture.

4.2) Hard Candies (Units in Weight-%)

components A* B* Trehalulose-containing compostion (ex. 1D) 92.34 92.34Reb A^(#) 0.06 0.06 Citric acid — 1.55 Herbal essence 0.78 — Flavouringagent 0.04 0.04 Flavouring agent (herbal flavour) 0.12 — Lactobionicacid 0.12 0.12 Colouring agent 0.66 0.01 Water 5.88 5.88 ex: example^(#)Reb A content >97 weight-% on dry matter of total Reb A sweetener*A: throat lozenge *B: lemon candy

4.3) Ice Cream (Units in Weight-%)

components Trehalulose-containing composition (ex. 1D) 22.17 Reb A^(#)0.04 Lactobionic acid 0.90 Skim milk (sugar content 53%) 8.07 butter(milk fat 82%) 10.09 Emulsifying agent 0.30 Stabilizer 0.22 Flavouringagent 0.20 Colouring agent 0.02 Milk fat 8.25 Milk solids 16.21 Water57.99 ^(#)≧95 weight-% of Reb A, rebaudioside B, rebaudioside C,dulcoside A, stevioside, rubusoside and steviolbioside on dry matter oftotal Reb A sweetener

4.4) Canned Orange (Units in Weight-%)

components Trehalulose-containing composition (ex. 1C) 11.00 Orange pulp67.80 Citric acid 0.40 Water 20.80

4.5) Carrot Juice (Units in Weight-%)

components Trehalulose-containing composition (ex. 1D) 10.70 Carrotjuice 30.00 Citric acid 0.05 Flavouring agent 0.10 Water 59.15

4.6) Fondant (Units in Weight-%)

Components Trehalulose-containing composition (ex. 1D) 97.8 Water 2.2

Example 5 Sweetening Power

To establish the relative sweetening power, the following solutions werecompared with one another in a triangle test with 15 testers in eachcase:

a) Two 7% strength sucrose solutions versus a 15.5% strength solution ofthe sweetener according to example 1D.b) Two 7% strength sucrose solutions versus a 17.5% strength solution ofthe sweetener according to example 1D.c) Two 7% strength sucrose solutions versus an 18.5% strength solutionof the sweetener according to example 1D.

In test a), six people identified the sweetener: no statisticallyverified difference from the sucrose solutions.

In test b), twelve people identified the present sweetener as “sweeter”:statistically verified difference with p=0.99.

In test c), likewise twelve people identified the present sweetener as“sweeter”: statistically verified difference with p=0.99.

The sweetening power of the sweetener according to the invention isabout half of that of sucrose. To increase the sweetening power, thesweetener can be mixed with for instance Reb A and lactobionic acid.

Example 6 Ice Cream

To produce ice cream with the sweetener of example 1D, 22.1 kg of dairycream (40% fat in dry matter), 58.1 kg of whole milk (3.7% fat in DM)and 4.5 kg of skim milk powder were mixed with 15 kg of the sweetener ofexample 1D and 0.3 kg of stabilizer, homogenized and sterilized.

After the sterilization, 53 g of finely ground methylphenylalanylaspartate were added to the ice composition, stirred,whipped and frozen. The product has the same sweetness and the sametaste as ice cream produced with 15 kg of sugar.

In the case of fruit ice cream it may be particularly advantageous todispense with additional sweeteners, such as Reb A, because thesweetener brings out the taste of the fruit considerable better.

Example 7 Strawberry jam

To produce a low-calorie strawberry jam, 1 kg of chopped strawberrieswas boiled together with 1 kg of the sweetener of example 1C and 8 g ofa medium-esterified pectin and 7 g of tartaric acid for three minutesand bottled in prepared bottles.

Comparison with a jam produced with sucrose showed no difference inconsistency, the sweetness was somewhat less but this was compensated bythe strawberry taste being detectably stronger. After storage for aperiod of six months, the sweetener showed no tendency to crystallize.

Example 8

The effect of trehalulose on the blood glucose response (glycemic index,GI) and the insulin response (insulinemic index, II) was examined incomparison to isomaltulose.

The analysis was done as a randomized cross over analysis on ten healthysubjects of both sex.

Blood glucose- and insulin-profiles were taken after consumption ofisomaltulose or trehalulose (each 25 g carbohydrate and 250 ml water)for an interval of 0 to 180 minutes at the following time points 0, 15,30, 45, 60, 90, 120, 150 and 180 minutes.

The results are given in FIGS. 15 and 16. As the graph for the bloodglucose level for trehalulose shows, trehalulose provides an increasedand longer sustaining supply of glucose (GI), which is a supply longerand in sum greater than provided by isomaltulose.

The insulin response (II) of isomaltulose runs, as expected on the basisof the blood glucose response of isomaltulose, in parallel to the bloodglucose response. In contrast, trehalulose shows a significantly lowerinsulin response (II) than isomaltulose and a significantly lower bloodinsulin level in contrast to what has been expected by the blood glucoseresponse of trehalulose. Thus, trehalulose provides a longer, moresustained and greater blood glucose level than isomaltulose andsimultaneously provides unexpectedly a significantly lower insulinresponse compared to isomaltulose, which is also lower than expectedconsidering the blood glucose profile of trehalulose itself.

What is claimed is:
 1. A method for treating a glucose or insulinmetabolism-related condition or disease wherein vitalose(trehalulose-containing composition) is applied in a suitable amount toa subject in need thereof, thereby eliciting a low insulin response anda sustained release of glucose, and wherein the vitalose is prepared bya process wherein (a) a sucrose-containing composition is contactedunder appropriate conditions with cells or cell extracts frommicroorganisms of the genera Pseudomonas and Protaminobacter, and (b)the trehalulose-containing composition is produced.
 2. The method ofclaim 1, wherein the glucose or insulin metabolism-related disease isselected from the group consisting of diabetes, metabolism syndrome,glucose intolerance, insulin resistance, adipositas, overweight,obesity, hyperlipidemia, cancer, a liver disease, insulin sensitivityand arthroscleroses.
 3. The method of claim 1, wherein in the process ofpreparing the vitalose (trehalulose-containing composition) thesucrose-containing composition is dissolved or suspended in an aqueousmedium.
 4. The method of claim 1, wherein in the process of preparingthe vitalose (trehalulose-containing composition) the cells or the cellextracts are immobilized on at least one carrier.
 5. The method of claim1, wherein in the process of preparing the vitalose(trehalulose-containing composition) the cells or cell extracts of bothgenera of microorganisms are co-immobilized on the same carrier.
 6. Themethod of claim 1, wherein the vitalose (trehalulose-containingcomposition) comprises a trehalulose content of 20 to 95 weight-%, basedon the weight of dry matter.
 7. The method of claim 1, wherein thevitalose (trehalulose-containing composition) comprises an isomaltulosecontent of 8 to 50 weight-%, based on the weight of dry matter.
 8. Themethod of claim 1, wherein the vitalose (trehalulose-containingcomposition) is applied in a product that comprises said vitalose and atleast one additive.
 9. The method of claim 8, wherein the productcomprises 0.02 to 3.0 weight-% of the at least one additive and 97.0 to99.98 weight-% of the vitalose (trehalulose-containing composition),each based on the weight of dry matter.
 10. The method of claim 8,wherein the product comprises 3 to 95 weight-% of the vitalose(trehalulose-containing composition) and 5 to 97 weight-% of the atleast one additive, each based on the weight of dry matter.
 11. Themethod of claim 8, wherein the additive is at least one of a steviaextract and a steviolglycoside.
 12. The method of claim 8, wherein theadditive is lactobionic acid, lactobionic-δ-lacton, a salt oflactobionic acid or a mixture thereof.
 13. The method of claim 8,wherein the additive is selected from the group consisting of acidicflavors, fruit flavors, sweet flavors, savory flavors, salty flavors, ahigh intensity sweetener, a sugar alcohol, sucromalt, ribose, tagatose,trehalose, organic acid, fruit extract, a bulk sweetener, a fiber, aprebiotic agent, a thickener, a vitamin, a mineral, a preservative, afood color and a therapeutic agent.
 14. The method of claim 8, whereinthe additive is selected from the group consisting of creatine,polyphenole, L-carnitin, omega-3 polyunsaturated fatty acid, omega-6polyunsaturated fatty acid, green tea extract, epigallocatechingallate(EGCG), aminoacids and peptopro.
 15. The method of claim 8, wherein theadditive is a tryptophan-containing peptide composition.
 16. A methodfor treating a glucose or insulin metabolism-related condition ordisease wherein vitalose (trehalulose-containing composition) is appliedin a suitable amount to a subject in need thereof, thereby eliciting alow insulin response and a sustained release of glucose, and wherein thevitalose comprises 20 to 95 weight-% trehalulose, 8 to 50 weight-%isomaltulose and 0.1 to 2 weight-% glucose, each based on the weight ofdry matter.
 17. The method of claim 16, wherein the glucose or insulinmetabolism-related disease is selected from the group consisting ofdiabetes, metabolism syndrome, glucose intolerance, insulin resistance,adipositas, overweight, obesity, hyperlipidemia, cancer, a liverdisease, insulin sensitivity and arthroscleroses.
 18. The method ofclaim 16, wherein in the process of preparing the vitalose(trehalulose-containing composition) the sucrose-containing compositionis dissolved or suspended in an aqueous medium.
 19. The method of claim16, wherein in the process of preparing the vitalose(trehalulose-containing composition) the cells or the cell extracts areimmobilized on at least one carrier.
 20. The method of claim 16, whereinin the process of preparing the vitalose (trehalulose-containingcomposition) the cells or cell extracts of both genera of microorganismsare co-immobilized on the same carrier.
 21. The method of claim 16,wherein the vitalose (trehalulose-containing composition) comprises atrehalulose content of 20 to 95 weight-%, based on the weight of drymatter.
 22. The method of claim 16, wherein the vitalose(trehalulose-containing composition) comprises an isomaltulose contentof 8 to 50 weight-%, based on the weight of dry matter.
 23. The methodof claim 16, wherein the vitalose (trehalulose-containing composition)is applied in a product that comprises said vitalose and at least oneadditive.
 24. The method of claim 23, wherein the product comprises 0.02to 3.0 weight-% of the at least one additive and 97.0 to 99.98 weight-%of the vitalose (trehalulose-containing composition), each based on theweight of dry matter.
 25. The method of claim 23, wherein the productcomprises 3 to 95 weight-% of the vitalose (trehalulose-containingcomposition) and 5 to 97 weight-% of the at least one additive, eachbased on the weight of dry matter.
 26. The method of claim 23, whereinthe additive is at least one of a stevia extract and a steviolglycoside.27. The method of claim 23, wherein the additive is lactobionic acid,lactobionic-δ-lacton, a salt of lactobionic acid or a mixture thereof.28. The method of claim 23, wherein the additive is selected from thegroup consisting of acidic flavors, fruit flavors, sweet flavors, savoryflavors, salty flavors, a high intensity sweetener, a sugar alcohol,sucromalt, ribose, tagatose, trehalose, organic acid, fruit extract, abulk sweetener, a fiber, a prebiotic agent, a thickener, a vitamin, amineral, a preservative, a food color and a therapeutic agent.
 29. Themethod of claim 23, wherein the additive is selected from the groupconsisting of creatine, polyphenole, L-carnitin, omega-3 polyunsaturatedfatty acid, omega-6 polyunsaturated fatty acid, green tea extract,epigallocatechingallate (EGCG), aminoacids and peptopro.
 30. The methodof claim 23, wherein the additive is a tryptophan-containing peptidecomposition.